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Standarization of mouse and rat intestinal flora, and development of preservation and monitaring methods for the intestinal flora.

小鼠和大鼠肠道菌群的标准化，以及肠道菌群保存和监测方法的开发。

基本信息

批准号：
02558009
负责人：
MITSUOKA Tomotari
金额：
$ 11.65万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

The purpose of this project is first of all to produce of gnotobiotic mice and rats with normal physiological function for standardizing the intestinal flora of SPF mice and rats, and then to develop the preservation and monitoring system of standardized intestinal flora of mice and rats. Our results were as follows :1. Gnotobiotic (GB) rats with normal physiological function were produced by using the methods which already have established for production of gnotobiotic mice. Namely, 3 strains of Lactobachillus (L.acidophilus, L.murinus, L.salivarius) and 28 strains of Bacteroidaceae isolated from feces of conventional (CV) rats, and chloroform-treated CV rat feces (survive only clostridia) were inoculated into germfree (GF) rats. These ex-Gf rats were free from Clostridium difficile and eliminated Pseudomonas aeruginosa inoculated orally.2. Gb rat flora was kept in GF mice inoculated with fecal suspension of GB rats. The production of GB rats was possible only by oral inoculation of feces of GB rats and rat-flora-mice.3. Standardized mouse flora kept in the intestine of GB mice were inoculated into 4 different strains of GF mice (CF#1, IQI,NC,BALB/c). There were no differences in cecal size and the composition of fecal flora among GF mouse strains.4. In SPF rat colony established by transferring GB rats, composition of their fecal flora was kept stably in the colony.5. Preservation method for standardized mouse flora by freezing was examined. Finally, in 1 : 100 fecal suspension in nutrient broth with 10 or 20% glycerol kept in -80 C floral bacteria were kept for 1 year. By inoculation with preserved floral suspension into GF mice, GB mice with normal physiological function were able to be produced, and their floral composition was similar to original GB mice more than that kept in GB mouse intestine for 1 year.

本项目的目的是首先生产具有正常生理功能的无菌小鼠和大鼠，用于标准化SPF小鼠和大鼠的肠道植物群，然后开发标准化小鼠和大鼠肠道植物群的保存和监测系统。主要研究结果如下：1.通过使用已经建立的用于生产无菌小鼠的方法生产具有正常生理功能的无菌（GB）大鼠。即，将从常规（CV）大鼠粪便和氯仿处理的CV大鼠粪便（仅存活梭菌）中分离的3株乳杆菌（嗜酸乳杆菌、鼠乳杆菌、唾液乳杆菌）和28株拟杆菌科菌株接种到无菌（GF）大鼠中。这些ex-Gf大鼠均无艰难梭菌感染，经口接种的铜绿假单胞菌也被清除.将GB大鼠植物群保存在接种GB大鼠粪便悬液的GF小鼠中。经口接种GB大鼠粪便和鼠菌群小鼠粪便，才有可能产生GB大鼠.将保存在GB小鼠肠道中的标准化小鼠植物群接种到4个不同品系的GF小鼠（CF#1、IQI、NC、BALB/c）中。不同品系的GF小鼠盲肠大小和粪便植物群组成无差异.在转移GB大鼠建立的SPF鼠群中，其粪便植物群的组成保持稳定.研究了小鼠标准化植物群的冷冻保存方法。最后，在1：100的粪便悬浮液中，在含有10或20%甘油的营养肉汤中保持在-80 ℃下，将花状细菌保持1年。用保存的花悬液接种GF小鼠，可产生生理功能正常的GB小鼠，其花组成与原GB小鼠相似，比在GB小鼠肠道中保存1年的花组成更接近原GB小鼠。